scholarly journals The lectin level and the effect of abscisic acid on hemagglutinating activity during rye germination

2014 ◽  
Vol 58 (3) ◽  
pp. 343-350 ◽  
Author(s):  
Mirosława Ferens-Sieczkowska ◽  
Joanna Karczewska ◽  
Bronisława Morawiecka
Keyword(s):  
Abscisic Acid ◽  
Trace Amount ◽  
Protein A ◽  
Fold Increase ◽  
Hemagglutinating Activity ◽  
Lectin Activity ◽  
Immunological Properties ◽  
Early Germination ◽  
Molecular Masses

As we have previously found, the embryo is the only source of lectin in the mature, dry rye seed. During early germination the lectin activity decreased and most of it was found in the coleoptile. Leaves were found to contain only a trace amount of this protein. A 5-7 fold increase in the amount of lectin was found in grains which had imbibed for 6­12 hours in the presence of 10<sup>-5</sup>M and 10<sup>-4</sup>M ABA. At an ABA concentration of 10<sup>-4</sup>M, about 6 µg of lectin per germ was found even in grains which had been germinating for 5 days. The immunological properties and molecular masses of both RGA and RGA­like lectin accumulated in the presence of ABA were found to be identical.

scholarly journals Saccharomyces cerevisiae ubiquitin-like protein Rub1 conjugates to cullin proteins Rtt101 and Cul3 in vivo

2004 ◽  
Vol 377 (2) ◽  
pp. 459-467 ◽  
Author(s):  
Jose M. LAPLAZA ◽  
Magnolia BOSTICK ◽  
Derek T. SCHOLES ◽  
M. Joan CURCIO ◽  
Judy CALLIS
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein A ◽  
Fold Increase ◽  
Lysine Residue ◽  
Reading Frame ◽  
E3 Ligases ◽  
C Terminus ◽  
Dependent Modification ◽  
F Box Protein

In Saccharomyces cerevisiae, the ubiquitin-like protein Rub1p (related to ubiquitin 1 protein) covalently attaches to the cullin protein Cdc53p (cell division cycle 53 protein), a subunit of a class of ubiquitin E3 ligases named SCF (Skp1–Cdc53–F-box protein) complex. We identified Rtt101p (regulator of Ty transposition 101 protein, where Ty stands for transposon of yeast), initially found during a screen for proteins to confer retrotransposition suppression, and Cul3p (cullin 3 protein), a protein encoded by the previously uncharacterized open reading frame YGR003w, as two new in vivo targets for Rub1p conjugation. These proteins show significant identity with Cdc53p and, therefore, are cullin proteins. Modification of Cul3p is eliminated by deletion of the Rub1p pathway through disruption of either RUB1 or its activating enzyme ENR2/ULA1. The same disruptions in the Rub pathway decreased the percentage of total Rtt101p that is modified from approx. 60 to 30%. This suggests that Rtt101p has an additional RUB1- and ENR2-independent modification. All modified forms of Rtt101p and Cul3p were lost when a single lysine residue in a conserved region near the C-terminus was replaced by an arginine residue. These results suggest that this lysine residue is the site of Rub1p-dependent and -independent modifications in Rtt101p and of Rub1p-dependent modification in Cul3p. An rtt101Δ strain was hypersensitive to thiabendazole, isopropyl (N-3-chlorophenyl) carbamate and methyl methanesulphonate, but rub1Δ strains were not. Whereas rtt101Δ strains exhibited a 14-fold increase in Ty1 transposition, isogenic rub1Δ strains did not show statistically significant increases. Rtt101K791Rp, which cannot be modified, complemented for Rtt101p function in a transposition assay. Altogether, these results suggest that neither the RUB1-dependent nor the RUB1-independent form of Rtt101p is required for Rtt101p function. The identification of additional Rub1p targets in S. cerevisiae suggests an expanded role for Rub in this organism.

scholarly journals Rat lung lectin synthesis, degradation and activation. Developmental regulation and modulation by dexamethasone

1987 ◽  
Vol 245 (3) ◽  
pp. 683-690 ◽  
Author(s):  
L B Clerch ◽  
P L Whitney ◽  
D Massaro
Keyword(s):  
Binding Protein ◽  
Developmental Regulation ◽  
Fold Increase ◽  
Rat Lung ◽  
Absolute Rate ◽  
Lectin Activity ◽  
Developmentally Regulated ◽  
Rat Pups ◽  
The Absolute

Soluble lectins are widely distributed cell-agglutinating proteins. Their activity is developmentally regulated in several tissues, including the lung, but virtually nothing is known about the mechanisms of the developmental regulation or the turnover of these proteins. We studied mechanisms that might be responsible for the developmentally regulated changes in the activity of a lectin (beta-galactoside-binding protein) found in the lung, and determined if its activity or turnover could be modulated by treatment of rat pups with a glucocorticosteroid hormone (dexamethasone). Our studies on the activity and turnover of the lectin indicated that the peak of lectin activity (units/mg of protein) that occurred at age 12 days appeared to be brought about by two means: an increase in the activity of the lectin molecule itself (units/micrograms of lectin) that occurred at age 8 days, and 1.5-fold increase in the absolute rate of lectin synthesis at age 11 days. The decline in lectin activity was associated with a decrease in its rate of synthesis, return to the baseline extent of activation, and an increased rate of degradation. Treatment of rat pups with dexamethasone diminished the peak of lectin activity (units/mg of protein) by about 25%. This effect of dexamethasone was due, at least in part, to the complete prevention of activation of the lectin molecule (units/micrograms of lectin) and a premature increase in the rate of lectin degradation. Perhaps the normal fall in lectin activity after age 11 days is caused by mechanisms induced by the increase in serum corticosteroid that occurs at that age.

Expression, phosphorylation and nuclear localization of the human P1 protein, a homologue of the yeast Mcm 3 replication protein

1995 ◽  
Vol 108 (4) ◽  
pp. 1381-1389 ◽  
Author(s):  
D. Schulte ◽  
R. Burkhart ◽  
C. Musahl ◽  
B. Hu ◽  
C. Schlatterer ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Protein A ◽  
Fold Increase ◽  
S Phase ◽  
Human Protein ◽  
Replication Protein ◽  
High Sequence Similarity ◽  
Central Regions ◽  
Mcm Proteins ◽  
Carboxy Terminal

The human protein P1 belongs to a newly discovered class of mammalian nuclear proteins with high sequence homology to yeast replication proteins. We present the entire amino acid sequence of the human protein P1 as predicted from the cDNA sequence, and show that P1 shares three central regions of high sequence similarity (about 75%) and a highly hydrophilic carboxy-terminal region with the yeast Mcm3 replication protein. The human genome most probably contains one P1 gene which is activated when HeLa cells progress to S phase, as shown by a several-fold increase in P1-specific mRNA. However, the amounts of P1 protein do not detectably change during this period, but P1 protein becomes phosphorylated at the beginning of S phase. In contrast to the yeast Mcm proteins, which disappear from nuclei after initiation of DNA replication, protein P1 remains in the nucleus during and after S phase. P1 is dispersed in mitotic cells and may be excluded from binding to chromosomes.

scholarly journals 2712. Safety and Immunogenicity of two Doses of ExPEC4V Vaccine Against Extraintestinal Pathogenic Escherichia coli Disease in Healthy Adult Participants

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S954-S954
Author(s):  
William B Smith ◽  
Darren Abbanat ◽  
Bart Spiessens ◽  
Oscar Go ◽  
Wouter Haazen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Immune Responses ◽  
Geometric Mean ◽  
Protein A ◽  
Fold Increase ◽  
Vaccine Safety ◽  
Double Blind Study ◽  
Double Blind ◽  
Grade 3 ◽  
O Antigens

Abstract Background The ExPEC4V vaccine contains 4 Escherichia coli O-antigens (O1A, O2, O6A, O25B) conjugated to exotoxin protein A and is being studied for prevention of Invasive Extraintestinal pathogenic E. coli (ExPEC) Disease (IED). This phase-2 double-blind study assessed safety and immunogenicity of ExPEC4V Clinical Trial Material (CTM), manufactured via a redesigned process (optimized O1A strain). Methods Participants (≥18 years) in stable health were randomized (3:1) to receive ExPEC4V dose 4:4:4:8 μg PS/serotype or placebo on Day 1 and second vaccination on Day 181 (6 months after first vaccination). Participants will be followed for safety until end of study at Day 360. Reactogenicity and immunogenicity (by ELISA, opsonophagocytic killing [OPA] assays) were evaluated pre-vaccination, and 15 days after first and second vaccinations (Day 195). Results Of 100 participants randomized (mean age 56, 48% males) and vaccinated (ExPEC4V, n = 75; placebo, n = 25), 97 completed Day 30. Solicited local AEs were higher for ExPEC4V (38.7%) than placebo (20%); most frequent was pain/tenderness (38.7% vs 20%). Solicited systemic AEs were higher in ExPEC4V (49.3%) than placebo (20%); most frequent was fatigue (32% vs. 12%). No serious or grade 3 solicited local AEs were reported. One participant in ExPEC4V experienced a grade 3 solicited systemic fatigue considered vaccine-related by investigator. ExPEC4V demonstrated immune responses against all serotypes at Day 15. Geometric mean titer effective concentration rank by serotypes was O2 > O1A > O6 > O25B (Figures 1 and 2). At Day 15, ≥ 82% of participants in ExPEC4V and none in placebo had ≥2-fold increase from baseline of ELISA titer for all serotypes. In ExPEC4V, ≥47% had ≥2-fold increase from baseline of OPA titer for all serotypes, while 8% in placebo had ≥2-fold increase only for O6A. Good correlation was observed between ELISA and OPA across serotypes (r ≥ 0.76). Conclusion ExPEC4V elicited robust and functional immune responses across all serotypes and was well tolerated with no vaccine safety findings. This study supports the development of future multivalent ExPEC vaccine to prevent IED. Disclosures All authors: No reported disclosures.

scholarly journals Phagocytosis of phenylhydrazine oxidized and G-6-PD-deficient red blood cells: the role of cell-bound immunoglobulins

Blood ◽  
1991 ◽  
Vol 78 (7) ◽  
pp. 1818-1825 ◽  
Author(s):  
S Horn ◽  
N Bashan ◽  
J Gopas
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Protein A ◽  
Fluorescein Isothiocyanate ◽  
Fold Increase ◽  
Fc Receptor ◽  
Autologous Serum ◽  
Murine Macrophages

Abstract In this study, the role of Igs in the recognition and removal of oxidatively damaged human red blood cells (RBCs) was investigated. Phagocytosis of normal RBCs exposed to the oxidative hemolytic agent phenylhydrazine (Phz) and of glucose-6-phosphate dehydrogenase (G6PD)- deficient RBCs by murine macrophages was examined. A 40-fold increase in phagocytosis of RBCs treated with 3 mmol/L Phz was obtained both in the absence and presence of autologous serum, indicating that binding of autologous antibodies to the oxidized cells is not essential for phagocytosis. Yet, a basal number of IgG molecules was found to be present on the RBCs, as determined both by binding of 125I protein A and fluorescein isothiocyanate-antihuman Ig to the cells. Macrophage Fc receptors were found to be involved in the recognition of the RBCs, because phagocytosis was partially inhibited by incubating macrophages with bovine serum albumin (BSA) anti-BSA complexes, aIg (aggregated Igs), and anti-Fc receptor II monoclonal antibodies. Galactose/mannose inhibited phagocytosis of oxidized RBCs additively to aIg. Because phagocytosis was decreased when Phz-RBCs were incubated with F(ab')2 fragments of antihuman antibodies, it is suggested that the basal amount of Igs bound to the cells plays a role in the recognition of Phz- RBCs. G6PD-deficient RBCs were recognized and phagocytosed by murine macrophages without preexposure to oxidants in vitro (mean of 19 RBCs/100 macrophages). This phagocytosis was not affected by the addition of serum and was inhibited by incubating macrophages with galactose/mannose and the various Fc receptor blockers. A positive correlation between hemoglobin content and the number of cell-bound Igs to each patient erythrocytes was found. These results support the involvement of both an Fc and a lectin-like macrophage receptor in the recognition and phagocytosis of Phz-oxidized and G6PD-deficient RBCs and suggest opsonization as a possible physiologic process for the removal of severe damaged RBCs.

Abstract WP143: Identification of Socially-Regulated Circulating Microrna Signatures in Stroke

Stroke ◽  
2020 ◽  
Vol 51 (Suppl_1) ◽  
Author(s):  
Juneyoung Lee ◽  
Venugopal R Venna ◽  
Louise D McCullough
Keyword(s):  
Mirna Expression ◽  
Protein A ◽  
Fold Increase ◽  
Artery Occlusion ◽  
Mirna Biogenesis ◽  
Circulating Microrna ◽  
Key Enzyme ◽  
Socially Isolated ◽  
Underlying Mechanisms ◽  
Potential Biomarkers

Introduction: Epidemiological and clinical studies strongly support that social isolation (SI) is associated with higher mortality after stroke. SI has been identified as a risk factor in a wide variety of diseases, including stroke. However, few studies have attempted to identify the underlying mechanisms contributing to the detrimental effect of SI. To better understand systemic and global regulation networks governed by social factors, and in an attempt to identify viable biomarkers, we profiled changes in circulating microRNA (miRNA) expression in aged stroke mice after SI. Methods: Aged C57BL/6 mice (18-20 months) of both sexes were pair-housed (PH) or socially isolated (SI) for two weeks prior to sham or a 60-minute middle cerebral artery occlusion surgery. Mice were sacrificed two weeks after surgery and blood samples were obtained for profiling of plasma miRNAs. Using whole miRNAome analysis, differentially expressed miRNAs by isolation, stroke and sex differences were comprehensively identified and compared using bioinformatics. We further used mimic and antagomiR treatments to confirm a potential role of the identified miRNAs. Results: Stroke significantly changed 8 miRNAs including miR-467d-3p ( P =0.0014), miR-376b-3p ( P =0.013) and miR-297c-5p ( P =0.016) in PH groups (sham vs. stroke). In a comparison between PH vs. SI in stroke mice, stroke SI mice showed a differential miRNA profile (11 miRNAs) including let-7 family potentially targeting Dicer protein, a key enzyme for miRNA biogenesis. Interestingly, compared to sham SI mice, stroke SI mice had a 2-fold increase in the number of altered miRNAs (21 miRNAs) including miR-129-1-3p ( P =0.0075), miR-28a-3p ( P =0.0091) and miR-204-5p ( P =0.016). Finally, we found a strong sex difference in miRNA expression in response to SI. Conclusions: We identified several key miRNAs as potential biomarkers for 1) stroke, 2) SI and 3) SI/stroke. These miRNA signatures are highly unique and create a complex gene-regulatory network that depends on the social environment. Our data provide potential biomarkers and identify several miRNAs that could lead to significant improvements in diagnosis of elderly patients at elevated risk for the detrimental effects of loneliness.

scholarly journals Sucrose and Cytokinin Interactions in Relation to Ethylene and Abscisic Acid Production in the Regulation of Morphogenesis in Pelargonium × hortorum L.H. Bailey In Vitro

2015 ◽  
Vol 57 (1) ◽  
pp. 62-69 ◽  
Author(s):  
Agnieszka Wojtania ◽  
Elżbieta Węgrzynowicz-Lesiak ◽  
Michał Dziurka ◽  
Piotr Waligórski
Keyword(s):  
Abscisic Acid ◽  
Ethylene Production ◽  
Sucrose Concentration ◽  
Fold Increase ◽  
Shoot Formation ◽  
Inhibitory Effect ◽  
Synthesis Inhibitor ◽  
Subculture Period ◽  
Ethylene Action

Abstract The aim of the study was to determine the effect of exogenous sucrose and cytokinin on ethylene production and responsiveness in relation to the shoot formation of Pelargonium × hortorum ‘Bergpalais’ in vitro. Increasing the concentration of sucrose from 15 to 40 g L−1 in medium containing meta-topolin (mT) resulted in a two-fold decrease in the number of shoots and leaves as well as a reduction in ethylene production. The addition of ethylene synthesis inhibitor (AVG) to mT-medium significantly reduced the ethylene production and the shoot growth, but it had no significant influence on the shoot formation. The mT-induced shoot formation was, however, significantly reduced in the presence of ethylene action inhibitor (AgNO3), in a manner dependent on sucrose levels. At the end of the subculture period, increased sucrose concentrations (15–40 g L−1) in the presence of mT and AgNO3 resulted in a 3.7-fold increase in ethylene emission. At the same time, the supply of sucrose caused a 2.8-fold increase in the level of endogenous abscisic acid (ABA). Our results may suggest that the inhibitory effect of high sucrose concentration (30 and 40 g L−1) may depend on its influence on ethylene sensitivity. It also suggests that sucrose-regulation of the shoot formation of Pelargonium in vitro is mediated by ABA.

scholarly journals Both endocytic and endogenous protein degradation in fibroblasts is stimulated by serum/amino acid deprivation and inhibited by 3-methyladenine

1990 ◽  
Vol 272 (3) ◽  
pp. 577-581 ◽  
Author(s):  
K B Hendil ◽  
A M B Lauridsen ◽  
P O Seglen
Keyword(s):  
Amino Acid ◽  
Protein Degradation ◽  
Protein A ◽  
Fold Increase ◽  
Isolated Hepatocytes ◽  
Lysosomal Degradation ◽  
Endogenous Protein ◽  
Endocytic Protein ◽  
Deficient Medium ◽  
Epidermal Growth

Incubation of BHK-21 hamster fibroblasts in a serum- and amino acid-deficient medium caused a 3-fold increase in the degradation of endogenous protein, a doubling of the degradation of endocytosed epidermal growth factor, and an eightfold increase in the degradation of endocytosed alpha 2-macroglobulin. 3-Methyladenine (3MA) inhibited the deprivation-induced lysosomal degradation of both endogenous and endocytosed protein, but had no effect on basal (non-induced) degradation. 3MA also inhibited deprivation-induced protein degradation in human IMR-90 fibroblasts. Some inhibition of protein synthesis and of endocytic uptake of alpha 2-macroglobulin was observed in 3MA-treated BHK-21 cells, whereas cellular ATP levels were unaffected. These results are different from those obtained with isolated hepatocytes, and suggest that in some cells both endogenous and endocytic protein degradation may be accelerated as part of a general deprivation response.

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